EP3502650B1 - Verfahren von und vorrichtung zum aufbringen von flüssigkeit auf einem substrat - Google Patents

Verfahren von und vorrichtung zum aufbringen von flüssigkeit auf einem substrat Download PDF

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Publication number
EP3502650B1
EP3502650B1 EP17210499.4A EP17210499A EP3502650B1 EP 3502650 B1 EP3502650 B1 EP 3502650B1 EP 17210499 A EP17210499 A EP 17210499A EP 3502650 B1 EP3502650 B1 EP 3502650B1
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Prior art keywords
substrate
applicator surface
liquid
range
applicator
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EP17210499.4A
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English (en)
French (fr)
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EP3502650A1 (de
EP3502650C0 (de
Inventor
Franciscus Johannes Theresia Nijpels
Raimond Ravelli
Bart Wilhelmus Anna Maria Mathis BEULEN
Sanne THEWESSEM
Rene Jacobus Maria HENDERIKX
Pascal Paul Marie Huysmans
Carmen LÓPEZ-IGLESIAS
Jacobus Peter Johannes PETER
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Universiteit Maastricht
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Universiteit Maastricht
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Publication of EP3502650C0 publication Critical patent/EP3502650C0/de
Publication of EP3502650B1 publication Critical patent/EP3502650B1/de
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes
    • B01L3/0241—Drop counters; Drop formers
    • B01L3/0244—Drop counters; Drop formers using pins
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • B01L2200/147—Employing temperature sensors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/42—Low-temperature sample treatment, e.g. cryofixation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/2813—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on
    • G01N2001/282—Producing thin layers of samples on a substrate, e.g. smearing, spinning-on with mapping; Identification of areas; Spatial correlated pattern
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N2035/1027—General features of the devices
    • G01N2035/1034—Transferring microquantities of liquid
    • G01N2035/1037—Using surface tension, e.g. pins or wires
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/10—Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
    • G01N35/1009—Characterised by arrangements for controlling the aspiration or dispense of liquids
    • G01N35/1011—Control of the position or alignment of the transfer device

Definitions

  • the invention relates to a method of and an apparatus for applying liquid, in particular one or more samples, e.g. for imaging or diffraction experiments under cryogenic conditions, to a substrate, in particular a sample carrier, such as a film on a grid.
  • substrates include nanoreactors and nanofluidic devices, such as nanocapillary array membranes.
  • Samples include (single) particles, e.g. macromolecular compounds and/or complexes thereof, e.g. proteins, DNA, RNA, viruses, cells, bacteria, nanoparticles, and crystals, for imaging, e.g. in a cryo-electron microscope (EM), or diffraction experiments, e.g. inspection by ionizing radiation, under cryogenic conditions.
  • EM cryo-electron microscope
  • WO 2013/109405 , WO 2013/109406 and US2014360286 disclose methods for preparing an electron microscopy sample on an EM sample grid, comprising: dispensing a plurality of discrete specimens onto an EM sample grid in an ordered array of specimen locations, each specimen of the plurality of discrete specimens being placed into an individual specimen location in the array of locations.
  • Cryogenic sample-grid (cryo-EM grid) preparation and imaging methods ensure that biological specimens withstand the vacuum inside electron microscopes, allow their investigation while trapped at physiological conditions that conserve the structural arrangement of the biomolecules and reduce the effect of radiation damage.
  • Arnold continues: these preparation methods have not improved significantly over the last 20 years and have some major drawbacks: a 2-4 pL sample volume is required, and 99.9% of the sample volume is lost during grid preparation, due to an extensive blotting step made with filter paper. Furthermore, blotting with filter paper can lead to protein aggregation or denaturation.
  • the current state of the art sample preparation methods are recognized as one of the most significant limitations in cryo-EM.
  • Arnold relates to a method for cryo-electron microscopy (cryo-EM) that requires only 3-20 nL of sample to prepare a cryo-EM grid, depending on the protocol used. The sample is applied and spread on the grid by a microcapillary. This method is also described in WO 2017/005297 .
  • the method according to the present invention comprises the steps defined in claim 1.
  • the entire volume of the liquid is captured between the applicator surface and the substrate (including liquid that penetrates into or spreads over the substrate).
  • the applicator surface is moved further towards the substrate, thus increasing the width of the capillary bridge (relative to the direction of the movement of the applicator surface) and/or providing a supply of liquid for application within the capillary bridge.
  • the applicator surface is moved relative to the substrate in a (systematic) pattern, such as parallel straight lines or line segments (connected e.g. by turns), undulating lines or line segments, or a spiral, e.g. until the capillary bridge breaks.
  • a (systematic) pattern such as parallel straight lines or line segments (connected e.g. by turns), undulating lines or line segments, or a spiral, e.g. until the capillary bridge breaks.
  • the lines or segments of the same line partly overlap to form a wider layer.
  • the distance between the applicator surface and the substrate is reduced, e.g. the applicator surface is moved gradually or stepwise towards the substrate to compensate for the reduced volume of the liquid in the capillary bridge resulting from application of the liquid to the substrate.
  • the distance is preferably maintained in a range from 1 to 20 ⁇ m.
  • the liquid is applied in a pattern or layer having a thickness in a range from 10 nm to 10 ⁇ m, preferably in range from 10 to 200 nm, preferably in range from 10 to 150 nm.
  • Layer thickness can be influenced inter alia by the (adjustable) speed of the relative movement between the applicator surface and the substrate, the width of the applicator surface, the distance between the applicator surface and the substrate, which, at least with flat surfaces, defines the height of the capillary bridge, the viscosity of the liquid, and the (relative) hydrophilicity of the applicator surface and the substrate.
  • the applicator surface is defined at an end of a pin, in particular a solid pin (comprising no lumen or capillary).
  • the applicator surface is defined as at least one hydrophilic area, preferably a continuous hydrophilic area or an array of hydrophilic areas, in a hydrophobic area or surroundings and/or the applicator surface is continuous and flat or (slightly) concave or convex.
  • the applicator surface has an area in a range from 1,0.1E3 to 50.1E3 ⁇ m 2 , preferably in a range from 2,0.1E3 to 30.1E3 ⁇ m 2 .
  • the amount of liquid adhered to the applicator surface is smaller than 1000 pL (picoliter), preferably smaller than 500 pL, preferably smaller than 400 pL, preferably in a range from 10 to 300 pL, and/or the liquid on the applicator surface has a contact angle in a range from 20 to 90 degrees, preferably in a range from 25 to 55 degrees and/or the height of the liquid on the applicator surface is in a range from 1 to 30 ⁇ m.
  • These parameters can be influenced e.g. by selecting area ( ⁇ m 2 ), shape, and hydrophilicity of the applicator surface in dependence of the properties of the liquid.
  • Yet another embodiment comprises the step of determining the distance between applicator surface and the substrate, before the liquid touches the substrate, preferably before adhering an amount of the liquid to the applicator surface.
  • the distance can be determined e.g. by optics or by an inductive or an capacitive sensor.
  • the position of the applicator surface and/or of one or more locations on the substrate is measured, e.g. by positioning the tip of a pin defining the surface and one or more locations on the substrate in the focal plane of an on-axis camera, the measured position or positions are stored, preferably in a memory of a computer or controller memory, and the measured position or positions are used for positioning of the applicator surface relative to the substrate.
  • two or more different samples e.g. different substances and/or different concentrations of the same substance, are applied to the substrate.
  • At least one reservoir e.g. at least one pipette
  • at least one reservoir is provided to supply the applicator surface with sample or samples.
  • a droplet or meniscus is temporarily expelled from the reservoir, sample is taken from the droplet or meniscus by contacting the droplet or meniscus with the applicator surface or applicator surfaces, once or repeatedly until sufficient sample or samples has/have been applied to the substrate, and the droplet or meniscus is withdrawn into the reservoir.
  • the reservoir containing the sample can be stored for future use.
  • the substrate is exposed to a plasma, before applying the sample to the substrate.
  • the plasma will (glow) discharge the substrate rendering it hydrophilic, which in turn facilitates a substantially even application of the sample to the substrate.
  • the plasma cleans the substrate. It is preferred that the substrate is subjected to a plasma just prior to applying the sample to the substrate, e.g. less than 15 minutes, preferably less than 10 minutes before applying the sample to the substrate.
  • the temperature of the substrate and/or of the applicator surface is adjusted to a temperature around, at or above the dew point of the gas surrounding the substrate and/or the applicator surface, preferably to one or more tenths of a degree, e.g. in a range from 0,1 to 1,0 degrees, below or above the dew point temperature of the gas surrounding the substrate and/or the applicator surface.
  • the dew point temperature is calculated from temperature and humidity measurements of the gas surrounding the substrate and/or the applicator surface. The temperature of the substrate and/or the applicator surface is measured as well. These measurements are used in a closed feedback loop to control e.g. a Peltier element that thermally contacts the substrate and/or the applicator surface.
  • one or more of the above steps, preferably all steps are carried out in a closed chamber, preferably providing controlled temperature and humidity.
  • the invention also relates to an apparatus for applying liquid, in particular one or more samples, to a substrate according to claim 12.
  • the controller is configured to effect one or more of the steps of the method(s) described above.
  • the applicator surface is defined as at least one hydrophilic area, preferably a continuous hydrophilic area or an array of hydrophilic areas, in a hydrophobic area or surroundings and/or the applicator surface is continuous and flat or (slightly) concave or convex.
  • the applicator surface has an area in a range from 1,0.1E3 to 50.1E3 pm2, preferably in a range from 2,0.1E3 to 30.1E3 ⁇ m 2 .
  • the apparatus comprises an on-axis camera configured to determine the distance between applicator surface and a substrate in the substrate holder, in particular to measure the position of the applicator surface and/or of one or more locations on the substrate is measured, to store the measured position or positions, and to use the measured position or positions for positioning of the applicator surface relative to the substrate.
  • the substrate holder and the applicator surface are located inside a climate chamber.
  • the apparatus comprises a cooling and/or heating element for the substrate and/or the applicator surface, a temperature sensor and a humidity sensor, and a controller configured to calculate the dew point of the gas surrounding the substrate and/or the applicator surface and to adjust, by means of the element(s), the temperature of the substrate and/or the applicator surface to a temperature around, at or above the dew point of the gas surrounding the substrate and/or the applicator surface, preferably to within one or more tenths of a degree from the dew point.
  • the controller comprises a processor and a memory and is configured, e.g. programmed, to receive data on the positions of the substrate and the applicator surface and the temperature and humidity of the surrounding gas, to process such data, and to operate one or more drivers, e.g. stepper motors, that move the substrate and/or the applicator surface, and one or more cooling and/or heating elements.
  • the controller may be implemented in the form of any system including a processor and a memory that is capable of performing the functions described in this specification. Further, the controller may be coupled to one or more input/output (I/O) devices.
  • I/O input/output
  • Examples of input devices may include, but are not limited to, one or more camera's and/or sensors, one or more buttons, a (small) keyboard, touchscreen, or the like.
  • Examples of output devices may include, but are not limited to, a monitor or a display, speakers, or the like. Input and/or output devices may be coupled to the data processing system either directly or through intervening I/O controllers.
  • US 2002/083998 relates to deposit device, e.g. a pin, cooperating with a fluid source defines a precisely sized drop of fluid of small diameter on a drop carrying surface. By repeated action, minute drops of fluid can be deposited precisely in a dense array, preferably under computer control.
  • WO2008/034914 discloses a dispensing device wherein at least one of the distal tip and the substrate are moved laterally relative to the other in order to produce elongated test zones.
  • WO 03/000422 relates to a two-pin dispensing system comprising a pair of separately movable pins for holding a droplet of liquid therebetween. Each pin includes a tip spaced predetermined distance from the other pin to define a sample acquisition region.
  • WO 00/01798 elates to a ceramic tip and a random access print head for the transfer of microfluidic quantities of fluid.
  • the tip is used as a capillary or "gravity” pin to draw or collect source fluid and “spot” or deposit the fluid onto the target via physical contact (touch-off).
  • liquid includes substances comprising a liquid phase, such as liquids containing (large) proteins and, non-Newtonian fluids (such as pastes and blood), which substances are capable of forming a capillary bridge.
  • Typical samples are aqueous solutions of large biomolecules such as proteins.
  • samples such as fibres, vesicles, viruses, nanocrystals and polymers and any other solution or suspension of particulate sample could be applied to the grid by capillary coating.
  • Figures 1 to 7 show an example of method of applying liquid, such as one or more samples 1, to a substrate, such as a so-called EM-grid 2, by means of an applicator in the form of a metal pin 3, e.g. a tungsten pin having a diameter of 1.6 mm.
  • a metal pin 3 e.g. a tungsten pin having a diameter of 1.6 mm.
  • the pin 3 has a frusto-conical tip 4 with an (imaginary) top angle of approximately 20 degrees and defining a flat end surface with sharp edges, serving as an applicator surface 5.
  • the applicator surface was polished to obtain a platform having a diameter in a range from 50 to 200 pm, e.g. 100 ⁇ m.
  • the tip is ultrasonically cleaned in ethanol.
  • the applicator surface 5 at the end of the pin 3 is gradually positioned in the focal plane 6 of an on-axis camera 7 depicted as a lense.
  • the position at which the applicator surface is in focus is stored and used as a reference for later positioning of the pin 3 relative to the EM-grid 2.
  • the grid 2 is positioned (on a holder and brought) into the focal plane 6 of the on axis camera 7. Together with the position of the applicator surface, this provides the information needed to position the applicator surface at a defined distance away from the surface of the grid.
  • a focal series of the entire surface of the grid can be made in advance, to obtain a height map of the non-planar grid surface. This enables varying the absolute position of the pin (and thus the applicator surface) while maintaining a constant relative position of the pin to the grid.
  • the clean pin 3 is loaded with sample 1.
  • the sample is stored in a reservoir, in this example a pipette 8, from which a small (sub-microliter) mother droplet is temporarily aspirated, preferably inside a climate chamber.
  • the pin is brought into contact with the droplet, thus adhering an amount of the sample to the applicator surface of the pin.
  • the liquid on the applicator surface is visually inspected, shown in Figure 4 , and the contact angle between the liquid and the surface and the height of the liquid are determined.
  • Typical contact angles and heights observed on surfaces having a diameter in a range from 50 to 200 ⁇ m are in a range from 25 to 55 degrees and in range from 10 to 30 pm, respectively. Evaporation of the liquid can be mitigated using a dewpoint control feedback loop on the pin and/or the grid.
  • the pin 3 now loaded with a sub-nanoliter droplet, is moved towards (glow discharged) grid and until a capillary bridge 9 is formed, as shown in Figure 5 .
  • Suitable components for an apparatus for applying liquid to a substrate such as a holder for the substrate, a climate chamber, electrodes for generating a plasma, a visualization system, sensors, and a Peltier element, are known in the art and described in, e.g., PCT patent application PCT/EP2017/065466 .
  • the pin 3 is mounted in a frame 11 by means of resilient elements 12, allowing the pin to move resiliently in the axial direction.
  • resilient elements 12 By establishing a capillary bridge and retracting the frame away from the substrate 2 with a force smaller than the capillary force between the substrate and the applicator surface, the liquid holds the substrate and the pin together.
  • the pin will follow, within boundaries, the surface of the grid even if this surface is not flat.

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
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  • Pathology (AREA)
  • Application Of Or Painting With Fluid Materials (AREA)
  • Sampling And Sample Adjustment (AREA)

Claims (14)

  1. Verfahren zum Aufbringen einer oder mehrerer Flüssigkeitsproben (1) für Abbildungs- oder Beugungsexperimente unter kryogenen Bedingungen auf ein Substrat (2), aufweisend die folgenden Schritte:
    Anhaftenlassen einer Menge der Flüssigkeit (1) an eine Applikatoroberfläche (5), die an einem Ende eines massiven Stifts (3), der kein Lumen und keine Kapillare aufweist, definiert ist,
    Bewegen der Applikatoroberfläche (5) in Richtung auf das Substrat (2), bis die Flüssigkeit das Substrat (2) berührt und eine Kapillarbrücke (9) zwischen der Applikatoroberfläche (5) und dem Substrat (2) bildet,
    gekennzeichnet durch
    Bewegen der Applikatoroberfläche (5) entlang des Substrats (2), wobei die relative Bewegung die Kapillarbrücke aufgrund einer Kapillarscherwechselwirkung verformen wird und eine Linie oder Schicht der Flüssigkeit auf das Substrat aufbringen wird.
  2. Verfahren nach Anspruch 1, wobei, nachdem die Flüssigkeit (1) das Substrat (2) berührt, das gesamte Volumen der Flüssigkeit (1) zwischen der Applikatoroberfläche (5) und dem Substrat (2) gefangen ist.
  3. Verfahren nach Anspruch 1 oder 2, wobei, nachdem die Flüssigkeit (1) das Substrat (2) berührt, die Applikatoroberfläche (5) weiter gegen das Substrat (2) bewegt wird.
  4. Verfahren nach einem der vorstehenden Ansprüche, wobei die Applikatoroberfläche (5) in einem Muster relativ zu dem Substrat (2) bewegt wird.
  5. Verfahren nach einem der vorstehenden Ansprüche, wobei während des Aufbringens der Flüssigkeit (1) auf das Substrat (2) der Abstand zwischen der Applikatoroberfläche (5) und dem Substrat (2) verringert wird und/oder wobei dieser Abstand in einem Bereich von 1 bis 20 µm beibehalten wird, und/oder wobei die Flüssigkeit (1) in einem Muster oder einer Schicht mit einer Dicke im Bereich von 10 nm bis 10 pm, vorzugsweise in einem Bereich von 10 bis 200 nm, aufgebracht wird.
  6. Verfahren nach einem der vorstehenden Ansprüche, wobei die an der Applikatoroberfläche (5) anhaftende Menge Flüssigkeit (1) kleiner als 1000 pL (Pikoliter), vorzugsweise kleiner als 500 pL, vorzugsweise kleiner als 400 pL, vorzugsweise in einem Bereich von 10 bis 300 pL, ist und/oder wobei die Flüssigkeit (1) an der Applikatoroberfläche (5) einen Kontaktwinkel in einem Bereich von 20 bis 60 Grad, vorzugsweise in einem Bereich von 25 bis 55 Grad, hat und/oder die Höhe der Flüssigkeit in einem Bereich von 1 bis 30 µm ist.
  7. Verfahren nach einem der vorstehenden Ansprüche, aufweisend den Schritt eines Bestimmens des Abstands zwischen Applikatoroberfläche (5) und dem Substrat (2), bevor die Flüssigkeit (1) das Substrat (2) berührt, vorzugsweise vor Anhaftenlassen einer Menge der Flüssigkeit (1) an die Applikatoroberfläche (5).
  8. Verfahren nach Anspruch 7, wobei die Position der Applikatoroberfläche (5) und/oder eines oder mehrerer Orte an dem Substrat (2) gemessen wird, die gemessene Position oder gemessenen Positionen gespeichert werden und die gemessene Position oder gemessenen Positionen zum Positionieren der Applikatoroberfläche (5) relativ zu dem Substrat (2) verwendet werden.
  9. Verfahren nach einem der vorstehenden Ansprüche, wobei zwei oder mehr unterschiedliche Proben (1) auf das Substrat (2) aufgebracht werden.
  10. Verfahren nach einem der vorstehenden Ansprüche, wobei mindestens ein Reservoir (8) bereitgestellt ist, um die Applikatoroberfläche (5) mit einer Probe oder Proben (1) zu versehen.
  11. Verfahren nach einem der vorstehenden Ansprüche, wobei zumindest vor und vorzugsweise auch während und nach Aufbringen der Flüssigkeit (1) auf das Substrat (2) die Temperatur des Substrats und/oder der Applikatoroberfläche auf eine Temperatur innerhalb 1,0 Grad unter oder über dem Taupunkt des das Substrat und/oder die Applikatoroberfläche umgebenden Gases, vorzugsweise auf innerhalb eines oder mehr zehntel Grad vom Taupunkt, angepasst wird.
  12. Vorrichtung zum Aufbringen einer oder mehrerer Flüssigkeitsproben (1) für Abbildungs- oder Beugungsexperimente unter kryogenen Bedingungen auf ein Substrat (2), aufweisend
    einen Substrathalter,
    eine Applikatoroberfläche (5), um daran eine Menge der Flüssigkeit (1) anhaften zu lassen, wobei die Oberfläche (5) an dem Ende eines massiven Stifts (3), der kein Lumen und keine Kapillare aufweist, definiert ist,
    einen oder mehrere Aktuatoren zum Bewegen der Applikatoroberfläche (5) und des Substrathalters aufeinander zu und relativ zueinander, und
    eine Steuerung, die konfiguriert ist, den oder die Aktuatoren anzutreiben, um die Applikatoroberfläche (5) und den Substrathalter aufeinander zu und relativ zueinander zu bewegen, um eine Kapillarbrücke (9) zwischen der Applikatoroberfläche (5) und einem Substrat (2) in dem Substrathalter zu bilden und die Flüssigkeit (1) auf die Oberfläche (2) aufzubringen,
    dadurch gekennzeichnet, dass die Flüssigkeit (1) auf die Oberfläche (2) aufgebracht wird durch Bewegen der Applikatoroberfläche (5) entlang des Substrats (2), wobei die relative Bewegung die Kapillarbrücke aufgrund einer Kapillarscherwechselwirkung verformen wird und eine Linie oder Schicht der Flüssigkeit auf das Substrat aufbringen wird.
  13. Vorrichtung nach Anspruch 12, aufweisend eine Auf-Achse-Kamera (7), die konfiguriert ist, den Abstand zwischen Applikatoroberfläche (5) und einem Substrat (2) in dem Substrathalter zu bestimmen.
  14. Vorrichtung nach Anspruch 12 oder 13, wobei der Substrathalter und die Applikatoroberfläche (5) im Innern einer Klimakammer angeordnet sind.
EP17210499.4A 2017-12-22 2017-12-22 Verfahren von und vorrichtung zum aufbringen von flüssigkeit auf einem substrat Active EP3502650B1 (de)

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WO2000001798A2 (en) * 1998-07-07 2000-01-13 Cartesian Technologies, Inc. Tip design and random access array for microfluidic transfer

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